Splitting Emission Signals

In fluorescence microscopy, there are several options available for viewing multiple colors from a single sample. The primary benefit of splitting the emission signals using dichroic beamsplitters is the ability to view multiple colors simultaneously rather than sequentially. This technique of using dichroics to separate emission signals is standard in flow instruments but requires specific arrangements when using fluorescence microscopes.

When the emission signals are being collected with a non-pixelated detector, such as a photomultiplier tube (PMT), any dichroic with appropriate transmission and blocking characteristics is acceptable. For signals being imaged with a pixelated detector, like a CCD, there are significant flatness requirements to consider.